Potent 1,2,4-Triazino[5,6 b]indole-3-thioether Inhibitors of the Kanamycin Resistance Enzyme Eis from Mycobacterium tuberculosis

Huy X. Ngo; Keith D. Green; Chathurada S. Gajadeera; Melisa J. Willby; Selina Y.L. Holbrook; Caixia Hou; Atefeh Garzan; Abdelrahman S. Mayhoub; James E. Posey; Oleg V. Tsodikov; Sylvie Garneau-Tsodikova

doi:10.1021/acsinfecdis.8b00074

Potent 1,2,4-Triazino[5,6 b]indole-3-thioether Inhibitors of the Kanamycin Resistance Enzyme Eis from Mycobacterium tuberculosis

Huy X. Ngo, Keith D. Green, Chathurada S. Gajadeera, Melisa J. Willby, Selina Y.L. Holbrook, Caixia Hou, Atefeh Garzan, Abdelrahman S. Mayhoub, James E. Posey, Oleg V. Tsodikov, Sylvie Garneau-Tsodikova

Research output: Contribution to journal › Article › peer-review

21 Scopus citations

Abstract

A common cause of resistance to kanamycin (KAN) in tuberculosis is overexpression of the enhanced intracellular survival (Eis) protein. Eis is an acetyltransferase that multiacetylates KAN and other aminoglycosides, rendering them unable to bind the bacterial ribosome. By high-throughput screening, a series of substituted 1,2,4-triazino[5,6b]indole-3-thioether molecules were identified as effective Eis inhibitors. Herein, we purchased 17 and synthesized 22 new compounds, evaluated their potency, and characterized their steady-state kinetics. Four inhibitors were found not only to inhibit Eis in vitro, but also to act as adjuvants of KAN and partially restore KAN sensitivity in a Mycobacterium tuberculosis KAN-resistant strain in which Eis is upregulated. A crystal structure of Eis in complex with a potent inhibitor and CoA shows that the inhibitors bind in the aminoglycoside binding site snugly inserted into a hydrophobic cavity. These inhibitors will undergo preclinical development as novel KAN adjuvant therapies to treat KAN-resistant tuberculosis.

Original language	English
Pages (from-to)	1030-1040
Number of pages	11
Journal	ACS Infectious Diseases
Volume	4
Issue number	6
DOIs	https://doi.org/10.1021/acsinfecdis.8b00074
State	Published - Jun 8 2018

Bibliographical note

Funding Information:
This study was funded by a National Institutes of Health (NIH) Grant No. AI090048 (to S.G.-T.), a grant from the Firland Foundation (to S.G.-T.), and a grant from the Center for Chemical Genomics (CCG) at the University of Michigan (to S.G.-T.), as well as by startup funds from the College of Pharmacy at the University of Kentucky (to S.G.-T. and O.V.T). H.X.N. was in part supported by a Pharmaceutical Sciences Excellence in Graduate Achievement Fellowship from the College of Pharmacy at the University of Kentucky. We thank S. Vander Roest, M. Larsen, and P. Kirchhoff from the CCG at the University of Michigan for their help with HTS. We thank the staff of sector 22 (SER-CAT) of the Advanced Photon Source at the Argonne National Laboratories for their assistance with the remote X-ray diffraction data collection. The beamline use was supported, in part, by the Center for Structural Biology at the University of Kentucky. We thank Dr. Wenjing Chen for preliminary work on IC50 value determination. We thank Helen V. Waldschmidt and Matthew D. Demars for the help in the preliminary synthesis of a few molecules. Use of trade names is for identification only and does not constitute endorsement by the U.S. Department of Health and Human Services, the U.S. Public Health Service, or the CDC. The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the funding agencies.

Publisher Copyright:
© 2018 American Chemical Society.

Keywords

aminoglycoside resistance
antitubercular agent
combination therapy
high-throughput screen
structure-activity relationship (SAR)

ASJC Scopus subject areas

Infectious Diseases

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Ngo, H. X., Green, K. D., Gajadeera, C. S., Willby, M. J., Holbrook, S. Y. L., Hou, C., Garzan, A., Mayhoub, A. S., Posey, J. E., Tsodikov, O. V., & Garneau-Tsodikova, S. (2018). Potent 1,2,4-Triazino[5,6 b]indole-3-thioether Inhibitors of the Kanamycin Resistance Enzyme Eis from Mycobacterium tuberculosis. ACS Infectious Diseases, 4(6), 1030-1040. https://doi.org/10.1021/acsinfecdis.8b00074

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title = "Potent 1,2,4-Triazino[5,6 b]indole-3-thioether Inhibitors of the Kanamycin Resistance Enzyme Eis from Mycobacterium tuberculosis",

abstract = "A common cause of resistance to kanamycin (KAN) in tuberculosis is overexpression of the enhanced intracellular survival (Eis) protein. Eis is an acetyltransferase that multiacetylates KAN and other aminoglycosides, rendering them unable to bind the bacterial ribosome. By high-throughput screening, a series of substituted 1,2,4-triazino[5,6b]indole-3-thioether molecules were identified as effective Eis inhibitors. Herein, we purchased 17 and synthesized 22 new compounds, evaluated their potency, and characterized their steady-state kinetics. Four inhibitors were found not only to inhibit Eis in vitro, but also to act as adjuvants of KAN and partially restore KAN sensitivity in a Mycobacterium tuberculosis KAN-resistant strain in which Eis is upregulated. A crystal structure of Eis in complex with a potent inhibitor and CoA shows that the inhibitors bind in the aminoglycoside binding site snugly inserted into a hydrophobic cavity. These inhibitors will undergo preclinical development as novel KAN adjuvant therapies to treat KAN-resistant tuberculosis.",

keywords = "aminoglycoside resistance, antitubercular agent, combination therapy, high-throughput screen, structure-activity relationship (SAR)",

author = "Ngo, {Huy X.} and Green, {Keith D.} and Gajadeera, {Chathurada S.} and Willby, {Melisa J.} and Holbrook, {Selina Y.L.} and Caixia Hou and Atefeh Garzan and Mayhoub, {Abdelrahman S.} and Posey, {James E.} and Tsodikov, {Oleg V.} and Sylvie Garneau-Tsodikova",

note = "Funding Information: This study was funded by a National Institutes of Health (NIH) Grant No. AI090048 (to S.G.-T.), a grant from the Firland Foundation (to S.G.-T.), and a grant from the Center for Chemical Genomics (CCG) at the University of Michigan (to S.G.-T.), as well as by startup funds from the College of Pharmacy at the University of Kentucky (to S.G.-T. and O.V.T). H.X.N. was in part supported by a Pharmaceutical Sciences Excellence in Graduate Achievement Fellowship from the College of Pharmacy at the University of Kentucky. We thank S. Vander Roest, M. Larsen, and P. Kirchhoff from the CCG at the University of Michigan for their help with HTS. We thank the staff of sector 22 (SER-CAT) of the Advanced Photon Source at the Argonne National Laboratories for their assistance with the remote X-ray diffraction data collection. The beamline use was supported, in part, by the Center for Structural Biology at the University of Kentucky. We thank Dr. Wenjing Chen for preliminary work on IC50 value determination. We thank Helen V. Waldschmidt and Matthew D. Demars for the help in the preliminary synthesis of a few molecules. Use of trade names is for identification only and does not constitute endorsement by the U.S. Department of Health and Human Services, the U.S. Public Health Service, or the CDC. The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the funding agencies. Publisher Copyright: {\textcopyright} 2018 American Chemical Society.",

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doi = "10.1021/acsinfecdis.8b00074",

language = "English",

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Ngo, HX, Green, KD, Gajadeera, CS, Willby, MJ, Holbrook, SYL, Hou, C, Garzan, A, Mayhoub, AS, Posey, JE, Tsodikov, OV & Garneau-Tsodikova, S 2018, 'Potent 1,2,4-Triazino[5,6 b]indole-3-thioether Inhibitors of the Kanamycin Resistance Enzyme Eis from Mycobacterium tuberculosis', ACS Infectious Diseases, vol. 4, no. 6, pp. 1030-1040. https://doi.org/10.1021/acsinfecdis.8b00074

TY - JOUR

T1 - Potent 1,2,4-Triazino[5,6 b]indole-3-thioether Inhibitors of the Kanamycin Resistance Enzyme Eis from Mycobacterium tuberculosis

AU - Ngo, Huy X.

AU - Green, Keith D.

AU - Gajadeera, Chathurada S.

AU - Willby, Melisa J.

AU - Holbrook, Selina Y.L.

AU - Hou, Caixia

AU - Garzan, Atefeh

AU - Mayhoub, Abdelrahman S.

AU - Posey, James E.

AU - Tsodikov, Oleg V.

AU - Garneau-Tsodikova, Sylvie

N1 - Funding Information: This study was funded by a National Institutes of Health (NIH) Grant No. AI090048 (to S.G.-T.), a grant from the Firland Foundation (to S.G.-T.), and a grant from the Center for Chemical Genomics (CCG) at the University of Michigan (to S.G.-T.), as well as by startup funds from the College of Pharmacy at the University of Kentucky (to S.G.-T. and O.V.T). H.X.N. was in part supported by a Pharmaceutical Sciences Excellence in Graduate Achievement Fellowship from the College of Pharmacy at the University of Kentucky. We thank S. Vander Roest, M. Larsen, and P. Kirchhoff from the CCG at the University of Michigan for their help with HTS. We thank the staff of sector 22 (SER-CAT) of the Advanced Photon Source at the Argonne National Laboratories for their assistance with the remote X-ray diffraction data collection. The beamline use was supported, in part, by the Center for Structural Biology at the University of Kentucky. We thank Dr. Wenjing Chen for preliminary work on IC50 value determination. We thank Helen V. Waldschmidt and Matthew D. Demars for the help in the preliminary synthesis of a few molecules. Use of trade names is for identification only and does not constitute endorsement by the U.S. Department of Health and Human Services, the U.S. Public Health Service, or the CDC. The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the funding agencies. Publisher Copyright: © 2018 American Chemical Society.

PY - 2018/6/8

Y1 - 2018/6/8

N2 - A common cause of resistance to kanamycin (KAN) in tuberculosis is overexpression of the enhanced intracellular survival (Eis) protein. Eis is an acetyltransferase that multiacetylates KAN and other aminoglycosides, rendering them unable to bind the bacterial ribosome. By high-throughput screening, a series of substituted 1,2,4-triazino[5,6b]indole-3-thioether molecules were identified as effective Eis inhibitors. Herein, we purchased 17 and synthesized 22 new compounds, evaluated their potency, and characterized their steady-state kinetics. Four inhibitors were found not only to inhibit Eis in vitro, but also to act as adjuvants of KAN and partially restore KAN sensitivity in a Mycobacterium tuberculosis KAN-resistant strain in which Eis is upregulated. A crystal structure of Eis in complex with a potent inhibitor and CoA shows that the inhibitors bind in the aminoglycoside binding site snugly inserted into a hydrophobic cavity. These inhibitors will undergo preclinical development as novel KAN adjuvant therapies to treat KAN-resistant tuberculosis.

AB - A common cause of resistance to kanamycin (KAN) in tuberculosis is overexpression of the enhanced intracellular survival (Eis) protein. Eis is an acetyltransferase that multiacetylates KAN and other aminoglycosides, rendering them unable to bind the bacterial ribosome. By high-throughput screening, a series of substituted 1,2,4-triazino[5,6b]indole-3-thioether molecules were identified as effective Eis inhibitors. Herein, we purchased 17 and synthesized 22 new compounds, evaluated their potency, and characterized their steady-state kinetics. Four inhibitors were found not only to inhibit Eis in vitro, but also to act as adjuvants of KAN and partially restore KAN sensitivity in a Mycobacterium tuberculosis KAN-resistant strain in which Eis is upregulated. A crystal structure of Eis in complex with a potent inhibitor and CoA shows that the inhibitors bind in the aminoglycoside binding site snugly inserted into a hydrophobic cavity. These inhibitors will undergo preclinical development as novel KAN adjuvant therapies to treat KAN-resistant tuberculosis.

KW - aminoglycoside resistance

KW - antitubercular agent

KW - combination therapy

KW - high-throughput screen

KW - structure-activity relationship (SAR)

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UR - http://www.scopus.com/inward/citedby.url?scp=85048251988&partnerID=8YFLogxK

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JO - ACS Infectious Diseases

JF - ACS Infectious Diseases

IS - 6

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Potent 1,2,4-Triazino[5,6 b]indole-3-thioether Inhibitors of the Kanamycin Resistance Enzyme Eis from Mycobacterium tuberculosis

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

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